Using ex vivo organotypic cultures to investigate donor cell integration in the gut

Disruption of the enteric nervous system (ENS), the largest branch of the peripheral nervous system, impacts on critical intestinal functions such as motility, fluid exchange and gastric acid/hormone secretion. Unfortunately, therapeutic interventions to treat ENS defects, are mainly limited to surgical resection of the affected region. However, over the past decade there has been an increasing focus on stem cell-based therapies for treating disease. Recent studies from our group, and others, have highlighted the potential of ENS progenitor-based therapy, as a means of replacing neurons after in vivo transplantation to mouse colon. (Cooper et al, 2016. PLoS One. 2016; 11:e0147989; Cooper et al, 2017. Neurogastroenterol Motil. 2017 Jan;29(1):e12900; McCann et al, 2017. Nature Communications 3;8:15937; Frith TJ et al, 2019 bioRxiv doi: https://www.biorxiv.org/content/10.1101/819748v1).

However, the precise mechanisms by which donor cells integrate within recipient tissue remain unclear. Importantly, previous studies have relied heavily on in vivo surgical transplantation procedures to rodents. While this has provided proof-of-principal data that donor cells can integrate within organs after transplantation, technical limitations in tissue opacity and in vivo imaging have limited the mechanistic investigation of how donor cells integrate. Recently, we have developed an ex vivo organotypic culture method which allows for long-term culture of murine gut segments. Using this approach, it is possible to reduce animal usage by >84%, as each individual mouse can generate up to 6 cultured colonic segments, as well as providing refinement in experimental technique as in vivo surgical transplantation is not required.

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Project grant

Status:

Not yet active

Principal investigator

Dr Conor McCann

Institution

University College London

Grant reference number

NC/V001078/1

Award date:

Sep 2020 - Aug 2022

Grant amount

£308,514